Overexpression, purification, crystallization and preliminary X-ray analysis of CheY4 from Vibrio cholerae O395

作者：Biswas Maitree; Khamrui Susmita; Sen Udayaditya; Dasgupta Jhimli^*

来源：Acta Crystallographica Section F-Structural Biology and Crystallization Communications, 2011, 67: 1645-1648.

DOI：10.1107/S1744309111041972

• Vibrio cholerae
• motility
• colonization

Chemotaxis and motility greatly influence the infectivity of Vibrio cholerae, although the role of chemotaxis genes in V. cholerae pathogenesis is poorly understood. In contrast to the single copy of CheY found in Escherichia coli and Salmonella typhimurium, four CheYs (CheY1CheY4) are present in V. cholerae. While insertional disruption of the cheY4 gene results in decreased motility, insertional duplication of this gene increases motility and causes enhanced expression of the two major virulence genes. Additionally, cheY3/cheY4 influences the activation of the transcription factor NF-?B, which triggers the generation of acute inflammatory responses. V. cholerae CheY4 was cloned, overexpressed and purified by NiNTA affinity chromatography followed by gel filtration. Crystals of CheY4 grown in space group C2 diffracted to 1.67 angstrom resolution, with unit-cell parameters a = 94.4, b = 31.9, c = 32.6 angstrom, beta = 96.5 degrees, whereas crystals grown in space group P3221 diffracted to 1.9 angstrom resolution, with unit-cell parameters a = b = 56.104, c = 72.283 angstrom, gamma = 120 degrees.

• 出版日期2011-12